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Alexa Fluor® 647 Mouse Anti-Human CD148
Alexa Fluor® 647 Mouse Anti-Human CD148

Analysis of Human CD148 Expression

       Left Panel - Two-parameter flow cytometric analysis of CD148 expression on human peripheral blood leucocytes. Whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The cells were washed and then stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714, Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CD148 antibody (Cat. No. 564732; Right Plot). Flow cytometric contour plots showing the expression profile of CD148 (or Ig isotype control staining) versus Side Scattered-Light signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes.

       Right Panel - Two-color flow cytometric analysis of CD148 expression on human peripheral blood lymphocytes. Erythrocyte-lysed whole blood was similarly washed and then stained with FITC Mouse Anti-Human CD19 antibody (Cat. No. 340409) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CD148 antibody (Right Plot). Two-color contour plots showing the coexpressed levels of CD19 versus CD148 (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

       Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.

Analysis of Human CD148 Expression

       Left Panel - Two-parameter flow cytometric analysis of CD148 expression on human peripheral blood leucocytes. Whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The cells were washed and then stained with either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Cat. No. 557714, Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CD148 antibody (Cat. No. 564732; Right Plot). Flow cytometric contour plots showing the expression profile of CD148 (or Ig isotype control staining) versus Side Scattered-Light signals (SSC) were derived from gated events with the forward and side light-scatter characteristics of viable leucocytes.

       Right Panel - Two-color flow cytometric analysis of CD148 expression on human peripheral blood lymphocytes. Erythrocyte-lysed whole blood was similarly washed and then stained with FITC Mouse Anti-Human CD19 antibody (Cat. No. 340409) and either Alexa Fluor® 647 Mouse IgG1, κ Isotype Control (Left Plot) or Alexa Fluor® 647 Mouse Anti-Human CD148 antibody (Right Plot). Two-color contour plots showing the coexpressed levels of CD19 versus CD148 (or Ig isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes.

       Flow cytometric analysis was performed using a BD FACSCanto™ II Flow Cytometer System.

Product Details
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BD Pharmingen™
DEP-1; DEP1; HPTP eta; HPTP-η; PTPRJ; R-PTP-eta; R-PTP-J; SCC1; p260
Human (QC Testing)
Mouse BALB/c IgG1, κ
Activated human peripheral blood mononuclear cells
Flow cytometry (Routinely Tested)
5 µl/test
VI 6T-083, M4
AB_2738925
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.

Product Notices

  1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  6. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564732 Rev. 1
Antibody Details
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A3

The A3 monoclonal antibody specifically binds to CD148. CD148 is a receptor-type tyrosine phosphatase that belongs to the receptor class 3 subfamily of the protein-tyrosine phosphatase family. CD148 is also known as Density-enhanced phosphatase 1 (DEP-1), Human protein tyrosine phosphatase- η (HPTP-η/HPTP-eta), or Protein tyrosine phosphatase receptor type J (PTPRJ). CD148 is a highly glycosylated type I transmembrane protein that is widely expressed on different cell types including monocytes, granulocytes, dendritic cells, thymocytes, T cells, B cells, NK cells, fibroblasts, and platelets. As a protein tyrosine phosphatase with receptor function, CD148 plays a role in regulating the signaling activities of various phosphorylated cellular proteins including the PDGF Receptor, catenins and Src family kinases.  In the immune system, CD148 plays a role in regulating the activities of different cell types. For example, upon T cell activation, CD148 expression is upregulated by T cells. CD148 can subsequently dephosphorylate LAT and PLC-γ signaling proteins and thereby downregulate T cell signaling responses. Immobilized A3 antibody can reportedly augment the proliferation of anti-CD3 antibody-stimulated peripheral blood T cells in culture.

564732 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564732 Rev.1
Citations & References
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Development References (4)

  1. Baker JE, Majeti R, Tangye SG, Weiss A. Protein tyrosine phosphatase CD148-mediated inhibition of T-cell receptor signal transduction is associated with reduced LAT and phospholipase Cgamma1 phosphorylation. Mol Cell Biol. 2001; 21(7):2393-2403. (Clone-specific: Flow cytometry). View Reference
  2. Scraven B, Hegen M, Autschbach F, Gaya A, Schwarz C, Meuer SC. CD148 (p260 phosphatase) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:576-580.
  3. Tangye SG, Phillips JH, Lanier LL, de Vries JE, Aversa G. CD148: a receptor-type protein tyrosine phosphatase involved in the regulation of human T cell activation. J Immunol. 1998; 161(7):9249-9255. (Immunogen: (Co)-stimulation, Flow cytometry, Functional assay). View Reference
  4. Whiteford JR, Xian X, Chaussade C, Vanhaesebroeck B, Nourshargh S, Couchman JR. Syndecan-2 is a novel ligand for the protein tyrosine phosphatase receptor CD148. Mol Cell Biol. 2011; 22(19):3609-3624. (Clone-specific: Flow cytometry, Functional assay, Western blot). View Reference
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564732 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.